Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3632] to Calnexin - BSA and Azide free
  • Suitable for: WB, IP, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Calnexin antibody [EPR3632] - BSA and Azide free
    See all Calnexin primary antibodies

  • Description

    Rabbit monoclonal [EPR3632] to Calnexin - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    Recognizes ER membrane, mitochondria and cis-Golgi

  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1, HEK-293T, U-2 OS, MCF7, THP-1 and RAW 264.7 cell lysates. ICC/IF: HAP1 and human lung carcinoma A549 cells. IHC-P: Human tonsil tissue.

  • General notes

    Ab232433 is the carrier-free version of ab92573. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab232433 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Western blot - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)
    Western blot - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)
    All lanes : Anti-Calnexin antibody [EPR3632] (ab92573) at 1/20000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : CANX knockout HEK-293T cell lysate
    Lane 3 : U-2 OS cell lysate
    Lane 4 : MCF7 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 80 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab92573).

    Lanes 1 - 4: Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab92573 was shown to react with Calnexin in wild-type HEK-293T cells in Western blot with loss of signal observed in CANX knockout cell line ab255368 (CANX knockout cell lysate ab263805). Wild-type HEK-293T and CANX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab92573 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 20000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)
    Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)

    ab92573 staining Calnexin in wild-type HAP1 cells (top panel) and CANX knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92573 at 1/1000 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92573).

  • Western blot - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)
    Western blot - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)
    All lanes : Anti-Calnexin antibody [EPR3632] (ab92573) at 1/20000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : Calnexin knockout HAP1 cell lysate
    Lane 3 : THP-1 cell lysate
    Lane 4 : RAW 264.7 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 68 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab92573 was shown to specifically react with Calnexin when Calnexin knockout samples were used. Wild-type and Calnexin knockout samples were subjected to SDS-PAGE. ab92573 and ab8245 (loading control to GAPDH) were diluted at 1/20,000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92573).

  • Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)
    Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433) Image from Mitsuda S et al. FEBS Open Bio. 2014;4:229-39. Fig 9(B).; doi: 10.1016/j.fob.2014.02.009 with permission from Elsevier.

    Immunocytochemistry/Immunofluorescence analysis of human lung carcinoma A549 cells labelling Calnexin with ab92573. Cells were fixed with 4% PFA in PBS for 15 minutes washed three times with PBS, and then washed twice with 25 mM glycine–PBS. The cells were treated with cold methanol and washed three times with 0.3% Triton X-100 in PBS for permeabilization. The fixed cells were blocked with 5% BSA in PBS overnight. An Alexa Fluor® 594-conjugated anti-rabbit was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92573).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)

    Immunohistochemical analysis of paraffin embedded Human tonsil tissue using ab92573 at a 1/100 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92573).

  • Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)
    Anti-Calnexin antibody [EPR3632] - BSA and Azide free (ab232433)

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